The primary goal of our research program is directed towards an understanding of cell wall biosynthesis on the molecular level. The synthesis, assembly, and amplification of this structure is catalyzed by an integrated series of membrane-associated enzymes that are carefully regulated by the organism. Our research will contribute to an understanding of these enzymes, membrane events in peptidoglycan synthesis, membrane-wall interrelationships, and membrane teichoic acid biosynthesis. In addition, it is our goal to apply the results of our studies to the design of antibacterial agents that inhibit the assembly of cross-linked peptidoglycan. Six specific objectives will be undertaken for the coming year. (1) Role of the wall in stabilizing the organization of those enzymes that are involved in processing nascent peptidoglycan. (2) Reconstitution of membranes and walls that are simultaneously freeze-thawed. (3) Processing of nascent peptidoglycan by freeze-thawed membranes for additon to walls. (4) Mechanisms of penicillin and D-methionine actions on nascent peptidoglycan synthesis. (5) Specific microenvironments monitored by the dansyl reporter group during the course of peptidoglycan assembly. (6) Mechanism and D-alanine acylation of lipoteichoic acid: role of D-alanyl-lipophilic intermediates and D-alanine: membrane acceptor ligase. An investigation of these projects will contribute to our understanding of cell wall assembly and membrane chemistry that relates to this process. Our present objectives will provide insights into the events, regulations, and translocations that occur in the bacterial membrane for elaboration of this extracellular structure, the cell wall.